Search PubMed⌕ Search

Biomedical subjects

M A Miles

Publications and source records attributed to M A Miles.

160 records · Page 9Linked to original sources
PubMedDate not supplied

Human urine stimulates growth of Leishmania in vitro.

Enhancement of trypanosomatid metacyclogenesis by insect urine components led us to test the effect of human urine as a culture additive. The addition of 1-5% urine to Schneider's Drosophila medium containing 10% foetal calf serum enhanced the growth of 11 Leishmania strains representing 8 different taxonomic groups. Cell division was stimulated in cultures with non-dividing organisms. Peak cell density was increased, as was the efficiency with which L. donovani could be isolated from infected hamsters. Preliminary work suggested that the modified medium would be useful for field isolation of L. donovani and L. braziliensis. The active nutrients or growth factors are not known.

Animals↗
PubMedDate not supplied

Chloroquine-resistant Plasmodium falciparum isolates from the Sudan lack two mutations in the pfmdr1 gene thought to be associated with chloroquine resistance.

Isolates of Plasmodium falciparum from 3 areas of the Sudan were recovered from cryopreservation in London and their chloroquine sensitivity was determined in vitro. Chloroquine resistance was detected in 6/6 isolates from Khartoum, 1/4 from Sennar and 3/3 from Gadarif, indicating that resistance is spreading. All the isolates were sensitive to mefloquine. Studies using blood spots on glass fibre discs and the polymerase chain reaction did not detect two mutations in the pfmdr1 gene, thought to be correlated with chloroquine-resistance, in any of the isolates studied.

Amino Acid Sequence↗
PubMedDate not supplied

Diagnosis of Giardia duodenalis infection in Bangladeshi infants: faecal antigen capture ELISA.

An enzyme-linked immunosorbent assay (ELISA) to detect antigens of Giardia duodenalis in faeces was evaluated as a diagnostic tool by testing faecal samples collected during a cohort study of 229 infants living in an urban slum in Dhaka, Bangladesh. Faecal samples had been collected at enrollment, on a routine monthly basis, and repeatedly during episodes of diarrhoea and infection with Giardia, and a portion of all samples was frozen in saline. A direct smear of all had been examined by microscopy and again after concentrating cysts by ether sedimentation. A total of 2121 of the 4936 samples stored during the 22 months study were tested by the ELISA. After excluding non-specific binding, the sensitivity of the assay was 94.2% and the specificity was 98%. The presence of other parasites, including flagellated protozoa, was not linked to false positive ELISA results. There was a correlation between the number of Giardia cysts present and the ELISA optical density. Assuming that the ELISA is 100% sensitive, microscopy detected 92.4% of the infections detected by the ELISA.

Animals↗
PubMedDate not supplied

Metacyclogenesis of Leishmania (Viannia) braziliensis in vitro: evidence that lentil lectin is a marker of complement resistance and enhanced infectivity.

Axenic culture in modified Grace's medium was used to induce metacyclogenesis of Leishmania (Viannia) braziliensis in vitro. Morphological characteristics, lectin agglutination profiles, susceptibility to complement lysis, and infectivity in vivo were compared between metacyclic promastigotes and promastigotes in mid-log phase growth. Short, arrow-like promastigotes and round, oval promastigotes were defined as putative metacyclic forms on the basis of being highly motile and free swimming, with a small cell body and long flagellum. These forms increased during metacyclogenesis to > 80% whereas long-bodied, slender promastigotes and intermediate slender promastigotes declined progressively. Lentil lectin selectively agglutinated L. braziliensis after the induction of metacyclogenesis, whereas concanavalin A, wheat germ agglutinin and peanut agglutinin similarly agglutinated metacyclic promastigotes and mid-log phase promastigotes. Metacyclic promastigotes survived in 7.5%-20% human serum whereas mid-log phase promastigotes did not. Five hundred metacyclic promastigotes were highly infective to hamsters whereas 500 mid-log phase promastigotes rarely caused any lesion. Specific agglutination by lentil lectin should allow purification of metacyclic organisms for standardization of immunoprotection and challenge experiments.

Agglutination↗
PubMedDate not supplied

Chagas' disease in the Amazon basin: V. Periurban palms as habitats of Rhodnius robustus and Rhodnius pictipes--triatomine vectors of Chagas' disease.

Trypanosoma cruzi infected Rhodnius robustus and/or Rhodnius pictipes were commonly found, in large numbers, in the Brazilian Amazonian palms Maximiliana regia ("inajá"), Acrocomia sclerocarpa ("mucajá") and Orbignya speciosa ("babaçu"). The common opossum, Didelphis marsupialis, was the animal most frequently associated with triatomine infested palms. R. pictipes, frequently light-attracted into houses from palm trees, was the probable source of an acute case of Chagas' disease in the vicinity of Belém. It is considered that triatomine infested palms are likely to cause some cases of acute Chagas' disease in the States of Amazonas and Rondônia. Possible control methods are suggested.

Animals↗
PubMedDate not supplied

Analysis of genetic diversity of Trypanosoma cruzi: an application of riboprinting and gradient gel electrophoresis methods.

Analysis of restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S from Trypanosoma cruzi yields a typical 'riboprint' profile that can vary intraspecifically. A selection of 21 stocks of T. cruzi and three outgroup taxa: T. rangeli, T. conorhini and Leishmania braziliensis were analysed by riboprinting to assess divergence within and between taxa. T. rangeli, T. conorhini and L. braziliensis could be easily differentiated from each other and from T. cruzi. Phenetic analysis of PCR-RFLP profiles indicated that, with one or two exceptions, stocks of T. cruzi could be broadly partitioned into two groups that formally corresponded to T. cruzi I and T. cruzi II respectively. To test if ribosomal 18S sequences were homogeneous within each taxon, gradient gel electrophoresis methods were employed utilising either chemical or temperature gradients. Upon interpretation of the melting profiles of riboprints and a section of the 18S independently amplified by PCR, there would appear to be at least two divergent 18S types present within T. cruzi. Heterogeneity within copies of the ribosomal 18S within a single genome has therefore been demonstrated and interestingly, this dimorphic arrangement was also present in the outgroup taxa. Presumably the ancestral duplicative event that led to the divergent 18S types preceded that of speciation within this group. These divergent 18S paralogues may have, or had, different functional pressures or rates of molecular evolution. Whether or not these divergent types are equally transcriptionally active throughout the life cycle, remain to be assessed.

Animals↗
PubMedDate not supplied

Visceral leishmaniasis in Teresina, n. e. Brazil: towards a DNA probe kit and its adaptation to processing blood-contaminated samples.

The Lmet2 chemiluminescent DNA probe is a valuable tool for identifying parasites of the Leishmania donovani -complex in sand flies, dogs and human samples. Recent blood meals in sand flies or blood contamination of tissue samples inhibited probe sensitivity, whether radiolabelled or chemiluminescent detection systems were used. Treatment of membranes with protease before hybridisation restored positive signal. Alternatively samples could be lysed with protease and applied to membranes with a vacuum blotting apparatus. The Lmet2 protocol provides the basis for a DNA probe kit that is adaptable for use with a wide range of other probes.

Animals↗
PubMedDate not supplied

Preliminary observations on the diagnosis and transmissibility of canine visceral leishmaniasis in Teresina, n.e. Brazil.

A pilot group of 49 dogs and control groups from non-endemic areas were examined serologically for the presence of visceral leishmaniasis (VL) by direct agglutination test (DAT), indirect immunofluorescence (IFAT) enzyme-linked immunosorbent assay (ELISA) and DOT-ELISA. Results indicated that DAT is less sensitive than the other assays and that serology with filter paper blood samples is less sensitive than with serum. Promastigote infections were common in fed Lutzomyia longipalpis taken from a dog kennel inhabited by a dog carrying Leishmania chagasi. Colony-bred Lu. longipalpis readily acquired L. chagasi infection when fed on skin lesions of dogs naturally infected with L. chagasi: a small proportion of flies also became infected when fed on apparently normal skin. Widespread distribution of amastigotes in normal skin of asymptomatic animals was shown both by intensive microscopy and by probing skin biopsy samples with the Lmet2 L. donovani-complex specific DNA probe. It was demonstrated that an immunologically naive dog could be infected by a single experimentally infected sand fly. Abundant amastigotes present within the resultant lesion 22 days later were transmissible to sand flies but serology remained negative at least 45 days after the infective bite. Experimental transmission of canine VL by sand fly bite is a valuable approach for determining which diagnostic procedures are most sensitive, specific and suitable for field application in suburban households.

Agglutination Tests↗
PubMedDate not supplied

Demonstration of natural Leishmania infection in asymptomatic dogs in the absence of specific humoral immunity.

Asymptomatic dogs from a Kala-Azar endemic region were screened for infection status by parasitological, immunological and molecular techniques. Bone marrow was examined for the presence of parasites by NNN culture and by using the Lmet 2 DNA probe. All the samples were negative in culture but 24 of 41 were positive as determined by the probe. Cellular and humoral immunity were detected by T cell proliferation assays and IFAT respectively. Specific cellular and humoral immunity were found in 20 and 26 dogs respectively out of a total of 41 dogs examined. The vast majority of dogs with Leishmania-specific antibodies were found to be parasitologically positive using the DNA probe while almost half those that had demonstrable cellular immunity were apparently parasite free. The observation that dogs can develop cellular immunity following natural infection clearly indicates that there is a spectrum of canine leishmaniasis similar to that observed in the human disease. The prevalence of dog leishmanial infection must also be higher than was presumed.

Animals↗